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Isolation of Two Distinct Epithelial Cell Lines from a Single Feline Mammary Carcinoma with Different Tumorigenic Potential in Nude Mice and Expressing Different Levels of Epidermal Growth Factor Receptors¹

Departments of Veterinary Pathology [J. M. H. M. M., J. A. M. S., W. M.] and Molecular Cell Biology [J. B.], State University Utrecht, Yalelaan 1, P.O.B. 80158, 3508 TD, Utrecht; Division of Tumor Biology, Netherlands Cancer Institute (Antoni van Leeuwenhoek Huis), Pleamanlaan 121, 1066 CX, Amsterdam [J. M. H. M. M., E. S., R. v. d. B., J. H., W. M.]; and Department of Pathology [C. C.], University Leiden, Leiden, The Netherlands

From a single spontaneous feline mammary carcinoma, two subpopulations of epithelial tumor cells have been isolated. The variant cells were established as cell lines designated K248C and K248P. DNA ploidy analysis showed that the two cell lines represented cell populations already present in the original tumor. Chromosome analysis confirmed the feline origin of K248C and K248P and demonstrated that in addition to unique marker chromosomes characteristic for each cell line, both cell lines had several marker chromosomes in common. These data suggest that the two cell populations arose from a hypothetical single ancestor which diverged during tumor progression. The K248C and K248P cell lines differed from one another with respect to their tumorigenicity in athymic mice and epidermal growth factor (EGF) receptor content. The K248C cells were highly tumorigenic as indicated by a short latency period and high take rate. The K248P cells were poorly tumorigenic. Southern blot analysis revealed that the K248C cells contained an amplified EGF receptor gene that was accompanied by elevated levels of EGF receptor RNA and protein. The K248C cells were growth inhibited in vitro at EGF concentrations that stimulated growth of K248P cells. The amplification of the EGF receptor gene could be detected only in DNA derived from K248C cells at high passage numbers and not in DNA derived from the original tumor and K248C cells at low passage numbers. These data suggest that amplification of the EGF receptor gene occurred during establishment of the K248C cell line.

¹ E. S. was supported by Dutch Cancer Society Grant NK87-12.

² To whom requests for reprints should be addressed.

Received 11/19/91. Accepted 5/24/91.

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